25 replies to this topic

[sgt. rock]

After getting a decent spore print in the bottom of a 1/2 pint jar I put a pre-sterilized lid w/ 1 needle hole on the jar,after injecting about 1/3 - 1/2 of a jar of steril water into the jar I usualy tape the needle hole with regular black electrical tape (and each time before filling a syringe from the jar i shake the jar)
DO YOU THINK THAT POSSIBLY CONTAMINANTS STUCK TO THE BLACK TAPE (I take it directly from the roll of tape to the jar lid) MAY BE MIXING WITH MY SPORE SOLUTION WHEN I SHAKE THE JAR?
Should i switch to medical tape? (it allows air in and moisture out)
15 OUT OF 16 PF jars in my last batch were contaminated!HELP!

[the_other_chap]

I'd say the tape was an unlikely contamination source.

Was the jar well sterilised before you took the print?
If it was, then I'd suggest the contaminations came from the mushroom cap.

Sure you PCd your substrate jars for long enough?

[sgt. rock]

I'd say the tape was an unlikely contamination source.

Was the jar well sterilised before you took the print?
If it was, then I'd suggest the contaminations came from the mushroom cap.

Sure you PCd your substrate jars for long enough?

Yea,I pre-sterilised the jar in the PC plus 10 min in the oven to dry it out.
Ya know,it might have been the cap...I fruit my new PF cakes along side 3rd and 4th flush cakes which sometimes get a little moldy,THAT HAS TO BE IT! Do you sugest fruiting cakes that i intend to get a cap/print from seperate from older mor questionable cakes? I have 2 extra aquariums.
So black tape over jar lid holes on spore solution jar is o.k.? The sticky side of electrical tape strait off the roll is steril enough?
PCd jars for 1 hour and 10 min.
Thanks everybody for your help. I GREW only PF Classic strain for 5 or 6 yrs and just switched to EQs 6 months ago and havent had a good batch since.

[the_other_chap]

I fruit my new PF cakes along side 3rd and 4th flush cakes which sometimes get a little moldy,THAT HAS TO BE IT! Do you sugest fruiting cakes that i intend to get a cap/print from seperate from older mor questionable cakes?

I would. I always print from 1st flush shrooms that haven't been anywhere near old or contaminated cakes.

If I'm printing in a jar, I also suspend the shroom cap a few mm above the bottom of the jar (nylon thread through the lid hole) to prevent contact contaminations.

[sgt. rock]

I would. I always print from 1st flush shrooms that haven't been anywhere near old or contaminated cakes.

If I'm printing in a jar, I also suspend the shroom cap a few mm above the bottom of the jar (nylon thread through the lid hole) to prevent contact contaminations.

I make a small tin foil "cup" about 3 inches deep inside the jar & cut a quarter-sized hole in the bottom.I PC the jar w/ the foil cup already in it w/ a lid on loosley.When its time to print i remove the jar lid and quickly sit the cap inside the foil cup (the bottom of the foil cup is 1/2 way from bottom of jar) then quickly sit the lid back on the jar but I dont tighten it just sit it on top of the jar then sit a large glass upside down on top of the jar.
The cap was from a 2nd flush cake and it fruited in a 10 gal aquarium alongside 3rd and 4th flush cakes that sometimes get a little moldy.THATS WHERE I WENT WRONG! I went 5 years without buying a print or syringe till' this batch.

[the_other_chap]

Yep, once those contamination spores start getting around the terrarium, nothing is safe!
By the time you notice most contaminations, they've already started producing and spreading spores :(

[mad_messiah]

i didn't do prints yet, but i did 2 liqid culture jars with success (first time) i used the tek thet hip tought me on chat, i drilled a 4mm hole in the lid and glued it with silicone, then sterelized and inoculated through thet silicone barier, prob you should try the same for the prints?
peace
MM

[sgt. rock]

i didn't do prints yet, but i did 2 liqid culture jars with success (first time) i used the tek thet hip tought me on chat, i drilled a 4mm hole in the lid and glued it with silicone, then sterelized and inoculated through thet silicone barier, prob you should try the same for the prints?
peace
MM

Damn,its like a little self-sealing injection port.
the silacone dosent shrink or melt in the PC?

[mad_messiah]

no, i even didn't wait untill it dries, just put in pc while it was soft and it came out plretty solid, had no problems with injection

[drgonzo1969]

will tape over the lids on my half pint jars slow colonization?

[Dank Side Of The Shroom]

dont tape the lid or the innoculation holes. the cakes need the air exchange.

[shedthemonkey]

yes the tape would slow it down. You have a verm layer, right? I like using two layers of postoffice tyvek as a filter over the metal lid but under the metal ring. some do one or the other or polyfil in a central hole or two. but it needs to breathe without drying out...which is why you dont just leave a filter cap too...a couple quarter inch holes are enough. No tape is best.

[hydrodad]

Shed is exactly right. Tape iS bad JUJU

[dbzeroone]

I've read many, many teks here that involve inoculation of jars via holes in the lid covered with tape. Seems to me that the easiest and most sterile way to do this would be to insert the needle THROUGH the tape covering the holes, but every tek says to peel the tape back and then recover the hole.

This mainly applies to LC's because the verm layer makes taping holes on pf jars unnecessary.

All I can think of is that any mold / bacteria on the tape surface could be carried through by the needle. Is this a greater risk than lifting the tape? Or is there a much greater risk here that I'm not seeing?

[iamsmiley]

sticking a needle in the tape leaves a pretty big hole and risks more contams than lifting tape in flow hood/glove box semi-sterile enviroment

[python]

i used to use tape on my pf jars in the pc only....to ensure the H20 /mix was not changed........however tape is not needed and really just a lot of work........

usually u remove tape before innocing jars and leave it off as the dry verm layer keeps contams at bay......and still allows the necessary air exchange

[Lazlo]

I do the same thing Python. Never had contams from open holes in the lids.

[Hippie3]

he's not talking pf tek,
but liquid culture.


frankly it does not make any difference
and you're proly better off
like you said, db,
just injecting thru your tape
and covering hole with more tape.

[BuckarooBanzai]

You might also consider creating some silicone injection ports in your lids. Injection ports keep your jars sealed and are self-healing after you inject through them. If you use sealed jars, remember to leave enough air space in them to colonize completely or use a double tyvek filter for air exchange.

Also, realize that the sealed jars will come out of the PC containing a vacuum. You have to use the airport syringe to vent the vacuum, or the jar will suck your syringe dry after you inject into the jar. Don't think that holding the plunger back will keep the syringe from getting sucked dry. It won't.

Airports:
http://mycotopia.net...ghlight=airport

Constructing a silicone ported/tyvek vented lid is detailed in this thread:
http://mycotopia.net...&highlight=tale

[doobydoobydoo]

The reason for tape as I understand is to prevent water from dripping/dropping into the holes in the tops of the jars and adding to much extra moisture and wetting that verm layer. I never use tape, have not for a LONG time. I just put a foil square wrapped over the jar tops when I PC them, and nothing over the holes from then on. The verm layer acts as the contam barrier, and you want air exchange. Tape can slow the air exchanges.